Membrane Topology of the ADP/ATP Carrier from Mitochondria

  • G. Brandolin
  • G. J. M. Lauquin
  • R. Brasseur
  • P. V. Vignais
Conference paper
Part of the NATO ASI Series book series (volume 83)


The exchange between ATP generated by oxidative phosphorylation in mitochondria and extramitochondrial ADP is catalysed by a specific carrier protein located in the inner mitochondrial membrane. Although ADP/ATP transport is well characterized on physiological and kinetic grounds, its detailed mechanism at the molecular level remains unknown. Due to severe difficulties encountered in the crystallisation of membrane transport proteins, no direct structural information on the ADP/ATP carrier is presently available. However, through a number of indirect approaches applied to the membrane-bound ADP/ATP carrier, it has been possible to propose a schematic view of the topographical organization of its polypeptide chain within the mitochondrial membrane. These approaches were theoretical and experimental, including prediction of the secondary structure from the amino-acid sequence, probing of binding sites with photochemical derivatives of inhibitors and substrates, and localisation of hydrophilic restricted segments of the peptide chain on one face of the membrane or the other using proteolytic and immunological techniques.


Brown Adipose Tissue Beef Heart Mitochondrial Carrier CATR Binding Site Phosphate Carrier 
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  1. Block MR, Lauquin GJM, Vignais PV (1982) Interaction of 3’-O (1-naphthoyl) adenosine 5’-diphosphate, a fluorescent adenosine 5’-diphosphate analog, with the adenosine 5’-diphosphate/adenosine 5’-triphosphate carrier protein in the mitochondrial membrane. Biochemistry 21: 5451–5457PubMedCrossRefGoogle Scholar
  2. Block MR, Lauquin GJM, Vignais PV (1983) Use of 3’-O (1-naphthoyl) membrane-bound adenosine 5’-diphosphate/adenosine 5’-triphosphate carrier. Biochemistry 22: 2202–2208PubMedCrossRefGoogle Scholar
  3. Block MR, Vignais PV (1984) Substrate-site interactions in the membrane-bound adenine nucleotide carrier as disclosed by ADP and ATP analogs. Biochim Biophys Acta 767: 369–376PubMedCrossRefGoogle Scholar
  4. Bogner W, Aquila H (1986) ADP/ATP carrier: analysis of transmembrane folding using pyridoxal phosphate. Methods Enzymol 125: 650–658PubMedCrossRefGoogle Scholar
  5. Bogner W, Aquila H, Klingenberg M (1986) The transmembrane arrangement of ADP/ATP carrier as elucidated by the lysine reagent pyridoxal 5-phosphate. Eur J Biochem 165: 611–620CrossRefGoogle Scholar
  6. Boulay F, Lauquin GJM, Tsugita A, Vignais PV (1983) Photolabeling approach to the study of the topography of the atractyloside 5’-triphosphate carrier protein. Biochemistry 22: 477–484PubMedCrossRefGoogle Scholar
  7. Boulay F, Lauquin GJM, Vignais PV (1986) Localization of immunoreactive regions in the beef heart adenine nucleotide carrier using rabbit antisera against the carboxyatractyloside-liganded and the sodium dodecyl sulfate denatured carrier forms. Biochemistry 25: 7567–7571PubMedCrossRefGoogle Scholar
  8. Boulay F, Vignais PV (1984) Localization of the N-ethylmaleimide reactive cystein in beef heart mitochondrial ADP/ATP carrier protein. Biochemistry 23: 4807–4812PubMedCrossRefGoogle Scholar
  9. Brandolin G, Boulay F, Dalbon P, Vignais PV (1989) Orientation of the N-terminal region of the membrane-bound ADP/ATP carrier protein explored by antipeptide antibodies and an arginine-specific endoprotease. Evidence that the accessibility of the N-terminal residues depends on the conformational state of the carrier. Biochemistry 28: 1093–1100PubMedCrossRefGoogle Scholar
  10. Brandolin G, Dupont Y, Vignais PV (1981) Substrate-induced fluorescence of the isolated ADP/ATP carrier protein in solution. Biochem Biophys Res Commun 98: 28–35PubMedCrossRefGoogle Scholar
  11. Brandolin G, Dupont Y, Vignais PV (1982) Exploration of the nucleotide binding sites of the isoplated ADP/ATP carrier protein from beef heart mitochondria. II. Probing of the nucleotide sites by formycin triphosphate, a fluorescent transportable analogue of ATP. Biochemistry 21: 6348–6353PubMedCrossRefGoogle Scholar
  12. Brandolin G, Dupont Y, Vignais PV (1985) Substrate-induced modifications of the intrinsic fluorescence of the isolated adenine nucleotide carrier protein: demonstration of distinct conformational states. Biochemistry 24: 1991–1997PubMedCrossRefGoogle Scholar
  13. Brasseur R, Lins L, Ruysschaert JM, Brandolin G, Vignais PV (1992) submittedGoogle Scholar
  14. Brdiczka D, Schumacher D (1976) Iodination of peripheral mitochondrial membrane proteins in correlation to the functional state of the ADP/ATP carrier. Biochem Biophys Res Commun 73: 823–832PubMedCrossRefGoogle Scholar
  15. Buchanan BB, Eiermann W, Riccio P, Aquila H, Klingenberg M (1976) Antibody evidencefor different conformational states of ADP/ATP translocator protein isolated from mitochondria. Proc Natl Sci Acad USA 73: 280–2284Google Scholar
  16. Capobianco L, Brandolin G, Palmieri F (1991) Transmembrane topography of the mitochondrial phosphate carrier explored by peptide-specific antibodies and enzymatic digestion. Biochemistry 39: 4963–4969CrossRefGoogle Scholar
  17. Dalbon P, Brandolin G, Boulay F, Hoppe J, Vignais PV (1988) Mapping of the nucleotide-binding sites in the ADP/ATP carrier of beef heart mitochondria by photolabeling with 2-azido(α-32P)adenosine diphosphate. Biochemistry 27: 5141–5149PubMedCrossRefGoogle Scholar
  18. Dupont Y, Brandolin G, Vignais PV (1982) Exploration of the nucleotide binding sites of the isolated ADP/ATP carrier protein from beef heart mitochondria. I. Probing of the nucleotide sites by naphthoyl-ATP, a fluorescent non transportable analogue of ATP. Biochemistry 21: 6343–6347PubMedCrossRefGoogle Scholar
  19. Duyckaerts C, Sluse-Goffart CM, Fux JP, Sluse FE, Liebecq C (1980) Kinetic mechanism of the exchange catalyzed by the adenine nucleotide carrier. Eur J Biochem 106: 1–6PubMedCrossRefGoogle Scholar
  20. Eckerskorn C, Klingenberg M (1987) In the uncoupling protein from brown adipose tissue the C-terminus protrudes to the c-side of the membrane as shown by tryptic cleavage. FEBS Letters 226: 166–170PubMedCrossRefGoogle Scholar
  21. Klingenberg M (1985) The ADP/ATP carrier in mitochondrial membrane. In: Martonosi A (ed) The Enzymes of Mitochondrial Membranes: Membrane Transport, vol 4, John Wiley, New York pp 511–553Google Scholar
  22. Klingenberg M (1989) Molecular aspects of the adenine nucleotide carrier from mitochondria. Arch Biochem Biophys 270: 1–14PubMedCrossRefGoogle Scholar
  23. Klingenberg M, Riccio P, Aquila H (1978) Isolation of the ADP, ATP carrier as the carboxyatractylate complex from mitochondria. Biochim Biophys Acta 503: 193–210PubMedCrossRefGoogle Scholar
  24. Lodish HF (1988) Multi-spanning membrane proteins: how accurate are the models? Trends Biochem Sci 13: 332–334PubMedCrossRefGoogle Scholar
  25. Marty I, Brandolin G, Vignais PV (1991) Topography of the membrane-bound ADP/ATP carrier assessed by enzymatic proteolysis. Biochemistry 31: 4058–4065CrossRefGoogle Scholar
  26. Mayinger P, Winkler E, Klingenberg M (1989) The ADP/ATP carrier from yeast (AAC-2) is uniquely suited for the assignment of the binding center by photoaffinity labeling. FEBS Lett 244: 421–426PubMedCrossRefGoogle Scholar
  27. Runswick MJ, Powell SJ, Nyren P, Walker JE (1987) Sequence of the bovine mitochondrial phosphate carrier protein: structural relationship to ADP/ATP translocase and the brown fat mitochondria uncoupling protein. EMBO J 6: 1367–1373PubMedGoogle Scholar
  28. Runswick MJ, Walker JE, Bisaccia F, Iacobazzi V, Palmieri F. (1990) Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins. Biochemistry 29: 11033–11040PubMedCrossRefGoogle Scholar
  29. Saraste M, Walker JE (1982) Internal sequence repeats and the path of polypeptide in mitochondrial ADP/ATP translocase. FEBS Lett 144: 250–254PubMedCrossRefGoogle Scholar
  30. Vignais PV, Block MR, Boulay F, Brandolin G, Lauquin GJM (1985) Molecular aspects of structure-function relationships in mitochondrial adenine nucleotide carrier. In: Bengha G (ed) Structure and Properties of Cell Membranes, vol II, CRC Press, pp 139-179Google Scholar
  31. Winkler E. and Klingenberg, M (1992) Photoaffinity labeling of the nucleotide-binding site of the uncoupling protein from hamster brown adipose tissue. Eur J Biochem 203: 295–304PubMedCrossRefGoogle Scholar
  32. Wohlrab H, Bukusoglu C, Defoe H (1989) Recent studies on the mitochondrial phosphate transport protein (PTP) and on its relationship to the ADP/ATP translocase (AAC) and the uncoupling. In: Azzi A, Nalecz KA, Nalecz MJ, Wojtczak L (eds) Anion Carriers of Mitochondrial Membranes. Springer-Verlag, Berlin Heidelberg New York, pp 113–121.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1994

Authors and Affiliations

  • G. Brandolin
    • 1
  • G. J. M. Lauquin
    • 1
    • 2
  • R. Brasseur
    • 1
    • 3
  • P. V. Vignais
    • 1
  1. 1.DBMS/Biochimie (URA 1130 CNRS alliée à l’INSERM)Centre d’Etudes NucléairesGrenoble cedexFrance
  2. 2.IBC-CNRSBordeaux cedexFrance
  3. 3.Faculté des SciencesBruxellesBelgique

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